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. 2015 Sep 2;4(9):e54. doi: 10.1038/emi.2015.54

Figure 3.

Figure 3

B. burgdorferi EF-Tu is not surface-localized. (A) IFAs of fixed (top panels) or unfixed (bottom panels) wild-type B. burgdorferi strain B31-A3 probed with monoclonal antibodies directed against outer surface protein OspA, periplasmic protein FlaB, or EF-Tu as primary antibodies, and FITC-conjugated goat anti-mouse antibody as secondary antibodies. (B) Proteinase K protection assay of wild-type B. burgdorferi. Intact cells were incubated with proteinase K (200 μg/mL) for 1 h. Washed cells were then subjected immunoblotting using a mixture of antibodies against OspA, FlaB, and EF-Tu. (C) Proteinase K protection assay of B. burgdorferi overexpressing EF-Tu. Intact cells were incubated with proteinase K (25, 50, 100, or 200 μg/mL) for 1 h and then subjected to SDS-PAGE (left panel and black arrow indicates the observed size of B. burgdorferi EF-Tu) or immunoblotting using a mixture of antibodies against OspA, FlaB, and EF-Tu (right panel). Data are representative of four separate experiments.